Screening A. ventricosa populations for 2n gametes

Nikoloudakis, Nikolaos; Aissat, Assia; Katsiotis, Andreas

doi:10.1007/s10681-017-2107-x

Cited by 8 publications

(9 citation statements)

References 38 publications

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“…Indeed, the examination of the size range of the pollen produced by an individual is the most common method to detect unreduced pollen. This method has been used as indicator of 2n pollen presence in several species (Quinn et al, 1974; Ramanna, 1983; Sala et al, 1989; Orjeda et al, 1990; Bretagnolle, 2001; Crespel et al, 2006; Kovalsky and Solís Neffa, 2012; Nikoloudakis et al, 2018). In these studies, pollen grains with larger size have usually been considered as unreduced.…”

Section: Discussionmentioning

confidence: 99%

Polyploidy in Fruit Tree Crops of the Genus Annona (Annonaceae)

et al. 2019

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Genome duplication or polyploidy is one of the main factors of speciation in plants. It is especially frequent in hybrids and very valuable in many crops. The genus Annona belongs to the Annonaceae, a family that includes several fruit tree crops, such as cherimoya (Annona cherimola), sugar apple (Annona squamosa), their hybrid atemoya (A. cherimola × A. squamosa) or pawpaw (Asimina triloba). In this work, genome content was evaluated in several Annona species, A. triloba and atemoya. Surprisingly, while the hybrid atemoya has been reported as diploid, flow cytometry analysis of a progeny obtained from an interspecific cross between A. cherimola and A. squamosa showed an unusual ploidy variability that was also confirmed karyotype analysis. While the progeny from intraspecific crosses of A. cherimola showed polyploid genotypes that ranged from 2.5 to 33%, the hybrid atemoyas from the interspecific cross showed 35% of triploids from a total of 186 genotypes analyzed. With the aim of understanding the possible implications of the production of non-reduced gametes, pollen performance, pollen size and frequency distribution of pollen grains was quantified in the progeny of this cross and the parents. A large polymorphism in pollen grain size was found within the interspecific progeny with higher production of unreduced pollen in triploids (38%) than in diploids (29%). Moreover, using PCR amplification of selected microsatellite loci, while 13.7% of the pollen grains from the diploids showed two alleles, 41.28% of the grains from the triploids amplified two alleles and 5.63% showed up to three alleles. This suggests that the larger pollen grains could correspond to diploid and, in a lower frequency, to triploid pollen. Pollen performance was also affected with lower pollen germination in the hybrid triploids than in both diploid parents. The results confirm a higher percentage of polyploids in the interspecific cross, affecting pollen grain size and pollen performance. The occurrence of unreduced gametes in A. cherimola, A. squamosa and their interspecific progeny that may result in abnormalities of ploidy such as the triploids and tetraploids observed in this study, opens an interesting opportunity to study polyploidy in Annonaceae.

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Section: Discussionmentioning

confidence: 99%

Polyploidy in Fruit Tree Crops of the Genus Annona (Annonaceae)

et al. 2019

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“…The estimation of proportions of unreduced (2n) gametes was one of the first proposed applications for pollen FCM [46], although rigorous descriptions of methodology have only appeared in recent years [4,11]. Recently, there has been an increased interest in estimating rates of 2n gamete production in plants using FCM, and some potentially promising applications include the experimental study of genetic and environmental drivers of gametic non-reduction [16], screening of natural plant populations for variation in rates of 2n gametes [10,15] In pollen samples, the peaks of interest in estimating 2n gamete production will almost certainly include events other than single gametic nuclei, including doublets (aggregates), debris, and contaminating somatic tissue nuclei [11]. Estimates of 2n gamete production that do not address this issue should be considered highly suspect, as Kron and Husband [11] demonstrated that 2C events in trinucleate species may consist of more than 80% doublets.…”

Section: Unreduced (2n) Gametesmentioning

confidence: 99%

“…Chopping is commonly used, probably because it is the predominant method for other tissues, and can be quite effective, particularly with germinated pollen, in which the pollen tubes help to hold the pollen grains in a stable mass and nuclei within pollen tubes are now outside the lysis‐resistant pollen wall [3]. Filter bursting, in which pollen grains are gently pressed against a mesh filter, was shown to be more efficient than chopping in a wide range of species [4], and has started to be used more frequently [10, 13–17]. The use of buffers that cause osmotic bursting also works in some species, and can produce very clean, low debris samples [18], although pollen of some species is very resistant to bursting in this way.…”

Section: Identification and Isolation Of Structures Of Interestmentioning

confidence: 99%

“…The estimation of proportions of unreduced (2 n ) gametes was one of the first proposed applications for pollen FCM [46], although rigorous descriptions of methodology have only appeared in recent years [4, 11]. Recently, there has been an increased interest in estimating rates of 2 n gamete production in plants using FCM, and some potentially promising applications include the experimental study of genetic and environmental drivers of gametic non‐reduction [16], screening of natural plant populations for variation in rates of 2 n gametes [10, 15], and assessing the relative contribution of 2 n male gametes to the overall complexity of reproductive interactions in taxonomically intriguing plant groups, for example, facultative apomicts, mixed‐ploidy systems; [17, 47].…”

Section: Research Applications In Pollen Fcmmentioning

confidence: 99%

“…Filter bursting, in which pollen grains are gently pressed against a mesh filter, was shown to be more efficient than chopping in a wide range of species [4], and has started to be used more frequently [10,[13][14][15][16][17]. The use of buffers that cause osmotic bursting also works in some species, and can produce very clean, low debris samples [18], although pollen of some species is very resistant to bursting in this way.…”

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confidence: 99%

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Flow cytometric analysis of pollen and spores: An overview of applications and methodology

et al. 2021

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Pollen grains are the male gametophytes in a seed-plant life cycle. Their small, particulate nature and crucial role in plant reproduction have made them an attractive object of study using flow cytometry (FCM), with a wide range of applications existing in the literature. While methodological considerations for many of these overlap with those for other tissue types (e.g., general considerations for the measurement of nuclear DNA content), the relative complexity of pollen compared to single cells presents some unique challenges. We consider these here in the context of both the identification and isolation of pollen and its subunits, and the types of research applications. While the discussion here mostly concerns pollen, the general principles described here can be extended to apply to spores in ferns, lycophytes, and bryophytes. In addition to recommendations provided in more general studies, some recurring and notable issues related specifically to pollen and spores are highlighted.

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